人基因XAPC7在肺腺癌细胞及组织中的表达和定位

Expression and Cellular Localization of XAPC7 in Lung Adenocarcinoma Cell Line and Tissue

目的了解XAPC7在肺腺癌细胞和组织中的表达及定位情况,为后续的功能研究提供线索。方法首先采用Western-blot技术检测XAPC7在HBE135-E6E7正常支气管上皮细胞株、A549肺腺癌细胞株、NCI-H446小细胞肺癌细胞株中的表达;再用细胞免疫荧光技术,以HBE135-E6E7和A549细胞为研究对象,用鼠抗人XAPC7单克隆抗体分析该蛋白的细胞定位情况;最后利用免疫组织化学法随机检测34例肺腺癌及相应癌旁组织中XAPC7蛋白的定位及表达。结果 Western-blot显示:与HBE135-E6E7相比,XAPC7在NCI-H446中表达明显降低(P<0.05),在A549中表达有所增加,但差异无统计学意义(P>0.05)。激光共聚焦显微镜观察发现XAPC7在A549中主要定位于细胞浆,胞核中少见,而在HBE135-E6E7中除胞浆外,胞核中也有较多分布。免疫组化显示:XAPC7在肺腺癌和肺鳞癌组织细胞的胞浆和胞核中均有表达。在34例肺腺癌中,癌组织胞浆中XAPC7蛋白的表达明显高于癌旁组织胞浆的表达(Z=-4.341,P=0.000),而在癌组织与癌旁组织的胞核中XAPC7蛋白的表达差异无统计学意义(Z=-0.167,P=0.867)。另外,XAPC7蛋白的表达与年龄、性别、肿瘤大小、组织分化和有无淋巴结转移无关(P均>0.05)。结论 XAPC7在肺腺癌细胞株中有表达,主要定位于胞浆。在肺腺癌组织和癌旁组织胞浆中的表达差异有统计学意义,为后续研究打下了基础。

Objective To investigate the expression and cellular localization of XAPC7 in lung adenocarcinoma cell line and tissue,and to provide the evidence for further study. Methods Western blot was used to measure the expression of XAPC7 in normal bronchial epithelial cell line HBE135-E6E7,lung adenocarcinoma cell line A549 and small cell lung cancer cell line NCI-H446.The monoclonal antibody against XAPC7 was used to identify the cellular localization of XAPC7 in HBE135-E6E7 and A549 cell by immunofluorescence technique.In addition,immunohistochemical method was used to detect the expression and cellular localization of XAPC7 in 34 cases of lung adenocarcinoma tissue and corresponding cancer-adjacent tissues. Results Western blot analysis showed that XAPC7 expression was significantly decreased in NCI-H446 cell（P〈0.05） but increased in A549 cell as compared with HBE135-E6E7 cell,and no statistically significant difference was found（P〉0.05）.Laser scanning confocal microscope found that XAPC7 was mainly located in the cytoplasm of A549 cell and rarely in the nucleus.However,in addition to the cytoplasm,XAPC7 had more distribution in the nucleus of HBE135-E6E7 cell.Immunohistochemical method showed that XAPC7 was expressed in the cytoplasm and nucleus of adenocarcinoma and squamous cell carcinoma of lung.In lung adenocarcinoma, XAPC7 expression in the cytoplasm of cancer cells was significantly higher than that in the cytoplasm of adjacent lung tissue（Z=-4.341,P=0.000）,but in the nucleus it did not show similar differences（Z=-0.167,P=0.867）. In addition,our data showed that XAPC7 expression was not associated with age,gender,tumor size,differentiation and lymph node metastasis（all P〉0.05）. Conclusions XAPC7 is expressed in lung adenocarcinoma cell line,primarily located in the cytoplasm.There is a statistically significant difference in XAPC7 expression in the cytoplasm between lung adenocarcinoma tissue and adjacent lung tissue.The research lays the foundation for further study.